Lateral Entorhinal Cortex Suppresses Drift in Cortical Memory Representations.

Memory retrieval is thought to depend on the reinstatement of cortical memory representations guided by pattern completion processes in the hippocampus. The lateral entorhinal cortex (LEC) is one of the intermediary regions supporting hippocampal-cortical interactions and houses neurons that prospectively signal past events in a familiar environment. To investigate the functional relevance of the activity of the LEC for cortical reinstatement, we pharmacologically inhibited the LEC and examined its impact on the stability of ensemble firing patterns in one of the efferent targets of the LEC, the medial prefrontal cortex (mPFC). When male rats underwent multiple epochs of identical stimulus sequences in the same environment, the mPFC maintained a stable ensemble firing pattern across repetitions, particularly when the sequence included pairings of neutral and aversive stimuli. With LEC inhibition, the mPFC still formed an ensemble pattern that accurately captured stimuli and their associations within each epoch. However, LEC inhibition markedly disrupted its consistency across the epochs by decreasing the proportion of mPFC neurons that stably maintained firing selectivity for stimulus associations. Thus, the LEC stabilizes cortical representations of learned stimulus associations, thereby facilitating the recovery of the original memory trace without generating a new, redundant trace for familiar experiences. Failure of this process might underlie retrieval deficits in conditions associated with degeneration of the LEC, such as normal aging and Alzheimer's disease.SIGNIFICANCE STATEMENT To recall past events, the brain needs to reactivate the activity patterns that occurred during those events. However, such reinstatement of memory traces is not trivial because it goes against the natural tendency of the brain to restructure the activity patterns continuously. We found that dysfunction of a brain region called the LEC worsened the drift of the brain activity when rats repeatedly underwent the same events in the same room and made them behave as if they had never experienced these events before. Thus, the LEC stabilizes the brain activity to facilitate the recovery of the original memory trace. Failure of this process might underlie memory problems in elderly and Alzheimer's disease patients with the degenerated LEC.

Outcome-Locked Cholinergic Signaling Suppresses Prefrontal Encoding of Stimulus Associations.

Acetylcholine (ACh) is thought to control arousal, attention, and learning by slowly modulating cortical excitability and plasticity. Recent studies, however, discovered that cholinergic neurons emit precisely timed signals about the aversive outcome at millisecond precision. To investigate the functional relevance of such phasic cholinergic signaling, we manipulated and monitored cholinergic terminals in the mPFC while male mice associated a neutral conditioned stimulus (CS) with mildly aversive eyelid shock (US) over a short temporal gap. Optogenetic inhibition of cholinergic terminals during the US promoted the formation of the CS-US association. On the contrary, optogenetic excitation of cholinergic terminals during the US blocked the association formation. The bidirectional behavioral effects paralleled the corresponding change in the expression of an activity-regulated gene, c-Fos in the mPFC. In contrast, optogenetic inhibition of cholinergic terminals during the CS impaired associative learning, whereas their excitation had marginal effects. In parallel, photometric recording from cholinergic terminals in the mPFC revealed strong innate phasic responses to the US. With subsequent CS-US pairings, cholinergic terminals weakened the responses to the US while developing strong responses to the CS. The across-session changes in the CS- and US-evoked terminal responses were correlated with associative memory strength. These findings suggest that phasic cholinergic signaling in the mPFC exerts opposite effects on aversive associative learning depending on whether it is emitted by the outcome or the cue.SIGNIFICANCE STATEMENT Drugs compensating for the decline of acetylcholine (ACh) are used for cognitive impairment, such as Alzheimer's disease. However, their beneficial effects are limited, demanding new strategies based on better understandings of how ACh modulates cognition. Here, we report that by manipulating ACh signals in the mPFC, we can control the strength of aversive associative learning in mice. Specifically, the suppression of ACh signals during an aversive outcome facilitated its association with a preceding cue. In contrast, the suppression of ACh signals during the cue impaired learning. Considering that this paradigm depends on the brain regions affected in Alzheimer's disease, our findings indicate that precisely timed control of ACh signals is essential to refine ACh-based strategies for cognitive enhancement.

Prefrontal Neural Ensembles Develop Selective Code for Stimulus Associations within Minutes of Novel Experiences.

Prevailing theories posit that the hippocampus rapidly learns stimulus conjunctions during novel experiences, whereas the neocortex learns slowly through subsequent, off-line interaction with the hippocampus. Parallel evidence, however, shows that the medial prefrontal cortex (mPFC; a critical node of the neocortical network supporting long-term memory storage) undergoes rapid modifications of gene expression, synaptic structure, and physiology at the time of encoding. These observations, along with impaired learning with disrupted mPFC, suggest that mPFC neurons may exhibit rapid neural plasticity during novel experiences; however, direct empirical evidence is lacking. We extracellularly recorded action potentials of cells in the prelimbic region of the mPFC, while male rats received a sequence of stimulus presentations for the first time in life. Moment-to-moment tracking of neural ensemble firing patterns revealed that the prelimbic network activity exhibited an abrupt transition within 1 min after the first encounter of an aversive but not neutral stimulus. This network-level change was driven by ∼15% of neurons that immediately elevated their spontaneous firing rates (FRs) and developed firing responses to a neutral stimulus preceding the aversive stimulus within a few instances of their pairings. When a new sensory stimulus was paired with the same aversive stimulus, about half of these neurons generalized firing responses to the new stimulus association. Thus, prelimbic neurons are capable of rapidly forming ensemble codes for novel stimulus associations within minutes. This circuit property may enable the mPFC to rapidly detect and selectively encode the central content of novel experiences.SIGNIFICANCE STATEMENT During a new experience, a region of the brain, called the hippocampus, rapidly forms its memory and later instructs another region, called the neocortex, that stores its content. Consistent with this dominant view, cells in the neocortex gradually strengthen the selectivity for the memory content over weeks after novel experiences. However, we still do not know precisely when these cells begin to develop the selectivity. We found that neocortical cells were capable of forming the selectivity for ongoing events within a few minutes of new experiences. This finding provides support for an alternative view that the neocortex works with, but not follows, the hippocampus to form new memories.

Lateral entorhinal cortex supports the development of prefrontal network activity that bridges temporally discontiguous stimuli.

The lateral entorhinal cortex (LEC) is an essential component of the brain circuitry supporting long-term memory by serving as an interface between the hippocampus and neocortex. Dysfunction of the LEC affects sensory coding in the hippocampus, leading to a view that the LEC provides the hippocampus with highly processed sensory information. It remains unclear, however, how the LEC modulates neural processing in the neocortical regions. To address this point, we pharmacologically inactivated the LEC of male rats during a temporal associative learning task and examined its impact on local network activity in one of the LEC's efferent targets, the prelimbic region of the medial prefrontal cortex (mPFC). Rats were exposed to two neutral stimuli, one of which was paired with an aversive eyelid shock over a short temporal delay. The LEC inhibition reduced the expression of anticipatory blinking responses to the reinforced stimuli without increasing responses to nonreinforced stimuli. In control rats, both the reinforced and nonreinforced stimuli evoked a short-lived, wide-band increase in the prelimbic network activity. With learning, the initial increase of gamma-band activity started to extend into the interval between the reinforced neutral stimulus and the eyelid shock. LEC inhibition attenuated the learning-induced sustained activity, without affecting the initial transient activity. These results suggest that the integrity of LEC is necessary for the formation of temporal stimulus associations and its neural correlates in the mPFC. Given the minimal effects on the innate network responses to sensory stimuli, the LEC appears not to be the main source of sensory inputs to the mPFC; rather it may provide a framework that shapes the mPFC network response to behaviorally relevant cues.

Neurobiology of systems memory consolidation.

Psychological theories posit that the hippocampus rapidly forms associations among ongoing events as they unfold. During a subsequent maturation process, so-called systems memory consolidation, these associations are gradually stabilized within distributed neocortical circuits through close interactions between the hippocampus and neocortex. In the past 50 years, a major effort in neurobiological research has been directed towards translating these descriptive accounts into tangible, biological processes in the brain. Until the early 2000s, most studies exclusively focused on examining whether the hippocampus becomes unnecessary for memory retrieval once the memory has been consolidated. With recent methodological advances, however, the field shifted attention to several other theoretical accounts and began to uncover the genetic, physiological and structural underpinnings of systems memory consolidation at an unprecedented level of precision. Here I review these neurobiological findings in the past 15 years within a framework of six essential predictions extracted from the psychological theories. Genetic approaches have made it possible to tag neurons that were activated during memory encoding and investigate their physiological and genetic profiles as well as reactivation patterns during subsequent retrieval. In parallel, electrophysiological and imaging approaches detected signs of the gradual refinement of memory representations and its underlying hippocampal-neocortical dialogue in millisecond-resolved neural firing patterns, the inter-region coupling of neural activity, across-day stability of neural ensemble activity and functional connectome. This summary represents substantial progress in our understanding of neurobiological mechanisms of systems memory consolidation whilst also identifying several essential remaining questions for future investigations.

Distributed representations of temporal stimulus associations across regular-firing and fast-spiking neurons in rat medial prefrontal cortex.

The prefrontal cortex has been implicated in various cognitive processes, including working memory, executive control, decision making, and relational learning. One core computational requirement underlying all these processes is the integration of information across time. When rodents and rabbits associate two temporally discontiguous stimuli, some neurons in the medial prefrontal cortex (mPFC) change firing rates in response to the preceding stimulus and sustain the firing rate during the subsequent temporal interval. These firing patterns are thought to serve as a mechanism to buffer the previously presented stimuli and signal the upcoming stimuli; however, how these critical properties are distributed across different neuron types remains unknown. We investigated the firing selectivity of regular-firing, burst-firing, and fast-spiking neurons in the prelimbic region of the mPFC while rats associated two neutral conditioned stimuli (CS) with one aversive stimulus (US). Analyses of firing patterns of individual neurons and neuron ensembles revealed that regular-firing neurons maintained rich information about CS identity and CS-US contingency during intervals separating the CS and US. Moreover, they further strengthened the latter selectivity with repeated conditioning sessions over a month. The selectivity of burst-firing neurons for both stimulus features was weaker than that of regular-firing neurons, indicating the difference in task engagement between two subpopulations of putative excitatory neurons. In contrast, putative inhibitory, fast-spiking neurons showed a stronger selectivity for CS identity than for CS-US contingency, suggesting their potential role in sensory discrimination. These results reveal a fine-scaled functional organization in the prefrontal network supporting the formation of temporal stimulus associations.NEW & NOTEWORTHY To associate stimuli that occurred separately in time, the brain needs to bridge the temporal gap by maintaining what was presented and predicting what would follow. We show that in rat medial prefrontal cortex, the former function is associated with a subpopulation of putative inhibitory neurons, whereas the latter is supported by a subpopulation of putative excitatory neurons. Our results reveal a distinct contribution of these microcircuit components to neural representations of temporal stimulus associations.

Cholinergic Modulation of Frontoparietal Cortical Network Dynamics Supporting Supramodal Attention.

A critical function of attention is to support a state of readiness to enhance stimulus detection, independent of stimulus modality. The nucleus basalis magnocellularis (NBM) is the major source of the neurochemical acetylcholine (ACh) for frontoparietal cortical networks thought to support attention. We examined a potential supramodal role of ACh in a frontoparietal cortical attentional network supporting target detection. We recorded local field potentials (LFPs) in the prelimbic frontal cortex (PFC) and the posterior parietal cortex (PPC) to assess whether ACh contributed to a state of readiness to alert rats to an impending presentation of visual or olfactory targets in one of five locations. Twenty male Long-Evans rats underwent training and then lesions of the NBM using the selective cholinergic immunotoxin 192 IgG-saporin (0.3 µg/µl; ACh-NBM-lesion) to reduce cholinergic afferentation of the cortical mantle. Postsurgery, ACh-NBM-lesioned rats had less correct responses and more omissions than sham-lesioned rats, which changed parametrically as we increased the attentional demands of the task with decreased target duration. This parametric deficit was found equally for both sensory targets. Accurate detection of visual and olfactory targets was associated specifically with increased LFP coherence, in the beta range, between the PFC and PPC, and with increased beta power in the PPC before the target's appearance in sham-lesioned rats. Readiness-associated changes in brain activity and visual and olfactory target detection were attenuated in the ACh-NBM-lesioned group. Accordingly, ACh may support supramodal attention via modulating activity in a frontoparietal cortical network, orchestrating a state of readiness to enhance target detection.SIGNIFICANCE STATEMENT We examined whether the neurochemical acetylcholine (ACh) contributes to a state of readiness for target detection, by engaging frontoparietal cortical attentional networks independent of modality. We show that ACh supported alerting attention to an impending presentation of either visual or olfactory targets. Using local field potentials, enhanced stimulus detection was associated with an anticipatory increase in power in the beta oscillation range before the target's appearance within the posterior parietal cortex (PPC) as well as increased synchrony, also in beta, between the prefrontal cortex and PPC. These readiness-associated changes in brain activity and behavior were attenuated in rats with reduced cortical ACh. Thus, ACh may act, in a supramodal manner, to prepare frontoparietal cortical attentional networks for target detection.

Neural representations of time-linked memory.

Many cognitive processes, such as episodic memory and decision making, rely on the ability to form associations between two events that occur separately in time. The formation of such temporal associations depends on neural representations of three types of information: what has been presented (trace holding), what will follow (temporal expectation), and when the following event will occur (explicit timing). The present review seeks to link these representations with firing patterns of single neurons recorded while rodents and non-human primates associate stimuli, outcomes, and motor responses over time intervals. Across these studies, two distinct firing patterns were observed in the hippocampus, neocortex, and striatum: some neurons change firing rates during or shortly after the stimulus presentation and sustain the firing rate stably or sidlingly during the subsequent intervals (tonic firings). Other neurons transiently change firing rates during a specific moment within the time intervals (phasic firings), and as a group, they form a sequential firing pattern that covers the entire interval. Clever task designs used in some of these studies collectively provide evidence that both tonic and phasic firing responses represent trace holding, temporal expectation, and explicit timing. Subsequently, we applied machine-learning based classification approaches to the two firing patterns within the same dataset collected from rat medial prefrontal cortex during trace eyeblink conditioning. This quantitative analysis revealed that phasic-firing patterns showed greater selectivity for stimulus identity and temporal position than tonic-firing patterns. Our summary illuminates distributed neural representations of temporal association in the forebrain and generates several ideas for future investigations.

Parvalbumin and GAD65 interneuron inhibition in the ventral hippocampus induces distinct behavioral deficits relevant to schizophrenia.

Hyperactivity within the ventral hippocampus (vHPC) has been linked to both psychosis in humans and behavioral deficits in animal models of schizophrenia. A local decrease in GABA-mediated inhibition, particularly involving parvalbumin (PV)-expressing GABA neurons, has been proposed as a key mechanism underlying this hyperactive state. However, direct evidence is lacking for a causal role of vHPC GABA neurons in behaviors associated with schizophrenia. Here, we probed the behavioral function of two different but overlapping populations of vHPC GABA neurons that express either PV or GAD65 by selectively inhibiting these neurons with the pharmacogenetic neuromodulator hM4D. We show that acute inhibition of vHPC GABA neurons in adult mice results in behavioral changes relevant to schizophrenia. Inhibiting either PV or GAD65 neurons produced distinct behavioral deficits. Inhibition of PV neurons, affecting ∼80% of the PV neuron population, robustly impaired prepulse inhibition of the acoustic startle reflex (PPI), startle reactivity, and spontaneous alternation, but did not affect locomotor activity. In contrast, inhibiting a heterogeneous population of GAD65 neurons, affecting ∼40% of PV neurons and 65% of cholecystokinin neurons, increased spontaneous and amphetamine-induced locomotor activity and reduced spontaneous alternation, but did not alter PPI. Inhibition of PV or GAD65 neurons also produced distinct changes in network oscillatory activity in the vHPC in vivo. Together, these findings establish a causal role for vHPC GABA neurons in controlling behaviors relevant to schizophrenia and suggest a functional dissociation between the GABAergic mechanisms involved in hippocampal modulation of sensorimotor processes.

Cholinergic, but not NMDA, receptors in the lateral entorhinal cortex mediate acquisition in trace eyeblink conditioning.

Anatomical and electrophysiological studies collectively suggest that the entorhinal cortex consists of several subregions, each of which is involved in the processing of different types of information. Consistent with this idea, we previously reported that the dorsolateral portion of the entorhinal cortex (DLE), but not the caudomedial portion, is necessary for the expression of a memory association between temporally discontiguous stimuli in trace eyeblink conditioning (Morrissey et al. (2012) J Neurosci 32:5356-5361). The present study examined whether memory acquisition depends on the DLE and what types of local neurotransmitter mechanisms are involved in memory acquisition and expression. Male Long-Evans rats experienced trace eyeblink conditioning, in which an auditory conditioned stimulus (CS) was paired with a mildly aversive electric shock to the eyelid (US) with a stimulus-free interval of 500 ms. Immediately before the conditioning, the rats received a microinfusion of neuroreactive substances into the DLE. We found that reversible inactivation of the DLE with GABAA receptor agonist, muscimol impaired memory acquisition. Furthermore, blockade of local muscarinic acetylcholine receptors (mACh) with scopolamine retarded memory acquisition while blockade of local NMDA receptors with APV had no effect. Memory expression was not impaired by either type of receptor blocker. These results suggest that the DLE is necessary for memory acquisition, and that acquisition depends on the integrity of local mACh receptor-dependent firing modulation, but not NMDA receptor-dependent synaptic plasticity.

Diversity of mnemonic function within the entorhinal cortex: a meta-analysis of rodent behavioral studies.

The entorhinal cortex (EC) has been shown to be an integral piece of the hippocampal memory system. It sits in a unique position within the brain with strong, intricate, reciprocal connectivity with the hippocampus as well as a vast array of neocortical regions. Topographical patterns of afferent and efferent projections suggest that the EC can be divided into the medial and lateral regions, each of which can be further divided into dorsal, intermediate, and lateral bands. These EC sub-regions, with variable anatomical features, indicate a multifaceted role of the EC in memory processing. The present article reviews rodent behavioral studies which tested the effect of manipulation to EC sub-regions in several different memory paradigms. An analysis of the specific targets of EC manipulations reveals an important role of the caudomedial EC for spatial memory. In recognition memory paradigms, damage to the lateral EC impairs recognition of the combined information of objects, locations, and environmental contexts relevant to the content of an experience; whereas damage to medial EC preferentially impairs the recognition of the spatial arrangement of objects relevant to the spatial location of an experience. Fewer studies have examined the impact of EC manipulations on contextual memory and temporal associative memory, the results of which are fairly conflicting and possible confounds are explored. Our summary provides further support for the functional dissociation within the EC for learning and memory and generates several ideas for future investigations.

Activation patterns in superficial layers of neocortex change between experiences independent of behavior, environment, or the hippocampus.

Previous work suggests that activation patterns of neurons in superficial layers of the neocortex are more sensitive to spatial context than activation patterns in deep cortical layers. A possible source of this laminar difference is the distribution of contextual information to the superficial cortical layers carried by hippocampal efferents that travel through the entorhinal cortex and subiculum. To evaluate the role that the hippocampus plays in determining context sensitivity in superficial cortical layers, behavior-induced expression of the immediate early gene Arc was examined in hippocampus-lesioned and control rats after exposing them to 2 distinct contexts. Contrary to expectations, hippocampal lesions had no observable effect on Arc expression in any neocortical layer relative to controls. Furthermore, another group of intact animals was exposed to the same environment twice, to determine the reliability of Arc-expression patterns across identical contextual and behavioral episodes. Although this condition included no difference in external input between 2 epochs, the significant layer differences in Arc expression still remained. Thus, laminar differences in activation or plasticity patterns are not likely to arise from hippocampal sources or differences in external inputs, but are more likely to be an intrinsic property of the neocortex.

The medial prefrontal cortex leaves the hippocampus when it prepares for the future.

Our memories help us plan for the future. In some cases, we use memories to repeat the choices that led to preferable outcomes in the past. The success of these memory-guided decisions depends on close interactions between the hippocampus and medial prefrontal cortex. In other cases, we need to use our memories to deduce hidden connections between the present and past situations to decide the best choice of action based on the expected outcome. Our recent study investigated neural underpinnings of such inferential decisions by monitoring neural activity in the medial prefrontal cortex and hippocampus in rats. We identified several neural activity patterns indicating awake memory trace reactivation and restructuring of functional connectivity among multiple neurons. We also found that these patterns occurred concurrently with the ongoing hippocampal activity when rats recalled past events but not when they planned new adaptive actions. Here, we discussed how these computational properties might contribute to success in inferential decision-making and propose a working model on how the medial prefrontal cortex changes its interaction with the hippocampus depending on whether it reflects on the past or looks into the future.

Functional dissociation within the entorhinal cortex for memory retrieval of an association between temporally discontiguous stimuli.

Anatomical connectivity and single neuron coding suggest a segregation of information representation within lateral (LEC) and medial (MEC) portions of the entorhinal cortex, a brain region serving as the primary input/output of the hippocampus and maintaining widespread connections to many association cortices. The present study aimed to expand this idea by examining whether these two subregions differentially contribute to memory retrieval for an association between temporally discontiguous stimuli. We found that reversible inactivation of the LEC, but not the MEC, severely impaired the retrieval of the recently and remotely acquired memory in rat trace eyeblink conditioning, in which a stimulus-free interval was interposed between the conditioned and unconditioned stimulus. Conversely, inactivation of the LEC had no effect on retrieval in delay eyeblink conditioning, where two stimuli were presented without an interval. Therefore, the LEC, but not the MEC, plays a long-lasting role in the retrieval of a memory for an association between temporally discontiguous stimuli.

Coupling of prefrontal gamma amplitude and theta phase is strengthened in trace eyeblink conditioning.

Trace eyeblink conditioning requires animals to associate a neutral stimulus (CS) and an aversive periorbital shock (US) that occurred moment later. Acquisition in this conditioning depends on several forebrain regions including the hippocampus, medial prefrontal, and entorhinal cortices in addition to the cerebellum. Activities of single and population neurons in these regions show several patterns of change with the conditioning. For example, the power and synchronization of theta oscillations are correlated with the rate of acquisition. Yet, little is known about how neuronal oscillations at other frequency bands change with the conditioning. The present study examined changes in gamma oscillations, which are typically associated with spiking activity of individual cells. We found that after CS offset the power of gamma oscillations at 35-45 Hz fluctuated at about 7 Hz. This rhythmic fluctuation of gamma power was observed in all three regions and locked to local theta oscillations at 4-8 Hz. Furthermore, over the course of 10 days of acquisition sessions, the coupling of gamma power and theta phase became stronger in the medial prefrontal cortex while it did not change in the hippocampus or lateral entorhinal cortex. Neither theta nor gamma power in any of the regions significantly changed across sessions, rejecting a possibility that the observed increase in prefrontal theta-gamma coupling was secondary to an increase in theta or gamma power. The theta-gamma coupling between different regions did not significantly change across sessions. These results suggest that prefrontal gamma oscillations become more effectively coordinated with concurrent theta oscillations in trace eyeblink conditioning. This may result in a stronger impact of prefrontal neuronal firing responses to the CS on the processing in down-stream regions, such as the lateral entorhinal cortex and/or pontine nuclei.

The cortical structure of consolidated memory: a hypothesis on the role of the cingulate-entorhinal cortical connection.

Daily experiences are represented by networks of neurons distributed across the neocortex, bound together for rapid storage and later retrieval by the hippocampus. While the hippocampus is necessary for retrieving recent episode-based memory associations, over time, consolidation processes take place that enable many of these associations to be expressed independent of the hippocampus. It is generally thought that mechanisms of consolidation involve synaptic weight changes between cortical regions; or, in other words, the formation of "horizontal" cortico-cortical connections. Here, we review anatomical, behavioral, and physiological data which suggest that the connections in and between the entorhinal and cingulate cortices may be uniquely important for the long-term storage of memories that initially depend on the hippocampus. We propose that current theories of consolidation that divide memory into dual systems of hippocampus and neocortex might be improved by introducing a third, middle layer of entorhinal and cingulate allocortex, the synaptic weights within which are necessary and potentially sufficient for maintaining initially hippocampus-dependent associations over long time periods. This hypothesis makes a number of still untested predictions, and future experiments designed to address these will help to fill gaps in the current understanding of the cortical structure of consolidated memory.

Prefrontal neuronal ensembles link prior knowledge with novel actions during flexible action selection.

We make decisions based on currently perceivable information or an internal model of the environment. The medial prefrontal cortex (mPFC) and its interaction with the hippocampus have been implicated in the latter, model-based decision-making; however, the underlying computational properties remain incompletely understood. We have examined mPFC spiking and hippocampal oscillatory activity while rats flexibly select new actions using a known associative structure of environmental cues and outcomes. During action selection, the mPFC reinstates representations of the associative structure. These awake reactivation events are accompanied by synchronous firings among neurons coding the associative structure and those coding actions. Moreover, their functional coupling is strengthened upon the reactivation events leading to adaptive actions. In contrast, only cue-coding neurons improve functional coupling during hippocampal sharp wave ripples. Thus, the lack of direct experience disconnects the mPFC from the hippocampus to independently form self-organized neuronal ensemble dynamics linking prior knowledge with novel actions.

Neuronal ensemble dynamics in associative learning.

Associative learning restructures the activity of numerous neurons distributed across cortical and subcortical regions. Individual neurons change the rate or timing of spiking patterns in response to environmental stimuli as they become associated with salient outcomes. Recent large-scale activity monitoring in rodents has uncovered that these learning-related changes occur concertedly across groups of neurons within and between brain regions. These changes yield neuronal representations of learned associations in three types of ensemble dynamics: ensemble firing rates, multineuron coactivity, and sequential activity. Here, I review some of the most robust demonstrations of these dynamics in the rodent neocortex and hippocampus and discuss their potential function in memory encoding, consolidation, and retrieval.

Flexibility of memory for future-oriented cognition.

Memories of daily experiences contain incidental details unique to each experience as well as common latent patterns shared with others. Neural representations focusing on the latter aspect can be reinstated by similar new experiences even though their perceptual features do not match the original experiences perfectly. Such flexible memory use allows for faster learning and better decision-making in novel situations. Here, I review evidence from rodent and primate electrophysiological studies to discuss how memory flexibility is implemented in the spiking activity of neuronal ensembles. These findings uncovered innate and learned coding properties and their potential refinement during sleep that support flexible integration and application of memories for better future adaptation.

Prefrontal projections to the nucleus reuniens signal behavioral relevance of stimuli during associative learning.

The nucleus reuniens (RE) is necessary for memories dependent on the interaction between the medial prefrontal cortex (mPFC) and hippocampus (HPC). One example is trace eyeblink conditioning, in which the mPFC exhibits differential activity to neutral conditioned stimuli (CS) depending on their contingency with an aversive unconditioned stimulus (US). To test if this relevancy signal is routed to the RE, we photometrically recorded mPFC axon terminals within the RE and tracked their changes with learning. As a comparison, we measured prefrontal terminal activity in the mediodorsal thalamus (MD), which lacks connectivity with the HPC. In naïve male rats, prefrontal terminals within the RE were not strongly activated by tone or light. As the rats associated one of the stimuli (CS+) with the US, terminals gradually increased their response to the CS+ but not the other stimulus (CS-). In contrast, stimulus-evoked responses of prefrontal terminals within the MD were strong even before conditioning. They also became augmented only to the CS+ in the first conditioning session; however, the degree of activity differentiation did not improve with learning. These findings suggest that associative learning selectively increased mPFC output to the RE, signaling the behavioral relevance of sensory stimuli.